Methylene bisacrylamide

Metformin, PMSF, leupeptin and aprotinin were purchased from Sigma (St. Louis, MO, USA). Acrylamide, methylene bisAcrylamide, sodium dodecyl sulfonate (SDS), Tris base, ammonium persulfate and Tween-20 were obtained from Amresco (Solon, OH, USA). Fetal bovine serum and DMEM were purchased from HyClone (Logan, UT, USA). Antibodies against p62, phospho-AMPKα (Thr172), phospho-mTOR (Ser2448), mTOR, phospho-p70 S6 Kinase (Thr421/Ser424), p70 S6 Kinase and β-actin were obtained from Cell Signaling (Danvers, MA, USA). Antibodies against PARP1, Caspase-9 and Caspase-3 were purchased from Abcam (Cambridge, MA, USA). Antibody against microtubule-associated protein 1 light chain 3 II (LC3-II) was from GeneTex (Irvine, CA, USA). Anti-rabbit/mouse goat immunoglobulin G-horseradish peroxidase (HRP) secondary antibodies were obtained from Santa Cruz (Santa Cruz, CA, USA). PVDF membrane was purchased from Millipore (Billerica, MA, USA). ECL Plus immunoblotting detection kit was from Bio-Rad (Hercules, CA, USA). Other routine reagents were obtained from Zhongshan (Beijing, China).

SDS, ammonium persulfate, sucrose, acrylamide, methylene bis-acrylamide, mannitol, glycerol and glycine were purchased from Amresco, LLC. Diazoxide, Nycodenz^®, urea, thiourea, EDTA and tetramethylethylenediamine were obtained from Sigma-Aldrich (Merck KGaA). Protein quantification kits, immobilized pH gradient (IPG) strips, dithiothreitol (DTT), BIO-Lyte, CHAPS, agarose, bromophenol blue, β-mercaptoethanol, iodoacetamide and PVDF were acquired from Bio-Rad Laboratories, Inc. Anti-cytochrome c oxidase subunit IV (COX IV; cat. no. ab14744), anti-2-oxoglutarate dehydrogenase (OGDH; cat. no. ab137773), anti-NADH dehydrogenase (ubiquinone) flavoprotein 1 (NDUFV1; cat. no. ab203208) and anti-NADH-ubiquinone oxidoreductase 75 kDa subunit (NDUFS1; cat. no. ab169540) antibodies were purchased from Abcam. All reagents were of analytical grade.